Immunohistochemical analysis of paraffin embedded breast carcinoma, using ab36772 diluted 1:50. Left: Untreated Ab; Right: Ab pre-treated with synthesized phosphopeptide.
All lanes : Anti-MEK6 (phospho S207) antibody (ab36722) at 1/500 dilutionLane 1 : Extracts from untreated MDA-MB-435 cells Lane 2 : Extracts from UV treated MDA-MB-435 cells Lysates/proteins at 30 µg per lane.
ICC/IF image of ab36722 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab36722, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.